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Methyltransferase Assays

TRANSCREENER® EPIGEN Methyltransferase Assay

Detection of HMTs with the Transcreener Epigen Methyltransferase Assay

Detection of HMTs with the Transcreener Epigen Methyltransferase Assay.
Enzyme titrations of Set7/9, G9a, and Suv39H1 histone methyltransferases with SAM at the indicated concentrations and a Histone H3-derived peptide as acceptor substrate.

Principle of Methyltransferase Assay

Principle of Methyltransferase Assay

Methyltransferase Assays and Kits from BellBrook Labs accelerate drug discovery by allowing simple, universal, HTS-ready assaying of methyltransferase activity.

  • Methyltransferase Assays from BellBrook Labs accelerate drug discovery by allowing simple, universal, HTS-ready assaying of methyltransferase activity. The Transcreener EPIGEN MT Assay provides universal methyltransferase detection in an HTS-proven format. It combines the extensively validated Transcreener AMP/GMP Assay with coupling enzymes that convert the SAH produced in a Methyltransferase reaction to AMP.

Features

  • Universal: detects any methyltransferase using any substrate
  • Robust: Z' values greater than 0.7 with low substrate consumption
  • Homogenous format: simple mix-and-read format for HTS workflows
  • Sensitive: reduces enzyme usage and allows use of physiological SAM concentrations
  • >24 hr reagent and signal stability: high quality data in automated workflows at any scale

Description

  • The Transcreener EPIGEN Methyltransferase Assay is a universal biochemical HTS assay for enzymes that produce S-adenosylhomocysteine (SAH), including all enzymes within the histone (HMTs) and DNA (DNMTs) methyltransferase families. It combines the extensively validated Transcreener AMP2/GMP2 Assay, which relies on fluorescent immunodetection of AMP, with coupling enzymes that convert SAH to AMP. Enzyme activity is signaled by a decrease in fluorescence polarization as the bound tracer is displaced from the Transcreener AMP2/GMP2 Antibody. The assay is a simple mix-and-read format with two liquid addition steps. Methyltransferase (MT) enzyme reactions are first quenched with Stop Buffer and then the SAH Detection Mixture containing coupling enzymes, Transcreener AMP2/GMP2 antibody, and tracer is added. The assay provides excellent signal at low substrate conversion, with a Z’ ≥ 0.7 and ≥ 100 millipolarization shift (mP) under normal reaction conditions.

Methyltransferase Overview

  • Methyltranferases are a diverse family of enzymes that catalyze the transfer of a methyl group from S-adenosylmethionine (SAM or AdoMet), the second most common enzymatic cofactor after ATP, to amino, thiol, or hydroxyl groups of acceptor molecules generating S-adenosylhomocysteine (SAH) as a byproduct. Example acceptor substrates include endogenous and xenobiotic small molecules, proteins, DNA and RNA, and lipids.
  • Methyltransferases play a role in epigenetic regulation through methylation of histones at lysine and arginine residues and methylation of DNA at cytosines in hemi-methylated CpG sites. There are over 50 Protein Lysine Methyltransferases (PKMTs) in humans, at least 10 Protein Arginine Methyltransferases (PRMTs), and three DNA Methyltransferases (DNMTs). Together these enzymes play a critical role in the dynamic modification of chromatin, and they are increasingly being targeted for cancer and other diseases with an epigenetic component.

Methyltransferase Assay Methods

  • Assay methods that detect SAH formation have the advantage of providing universal detection of MT enzymes regardless of the acceptor substrate or the mix of methylated reaction products. Universal assay methods reduce drug candidate assay development costs by using one single set of assay reagents for all methyltransferase targets. Direct immunodetection of SAH would be advantageous as it would eliminate the potential for compound interference from coupling enzymes, however it requires an antibody that specifically binds SAH in the presence of excess SAM; ie, that differentiates on the basis of a single methyl group. There is one literature report of an FP-based methyltransferase assay using an anti-SAH antibody from a diagnostic assay kit for homocysteine, but its commercial availability is unknown.
  • In the absence of an available, direct assay for SAH detection, coupled enzyme assays have been developed. BellBrook Labs utilizes two coupling enzymes to convert the SAH to AMP, which can then be measured with a monoclonal antibody and tracer with negligible cross-reactivity with SAM. This assay format can detect both histone methyltransferases and DNA methyltransferases.

ORDER INFORMATION

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TRANSCREENER EPIGEN Methyltransferase Assay

Order# Description Amount Price Data Sheet
3017-1K Transcreener EPIGEN Methyltransferase Assay EA 900,00 PDF
3017-10K Transcreener EPIGEN Methyltransferase Assay EA 5400,00 PDF
3017-100K Transcreener EPIGEN Methyltransferase Assay EA on request PDF
  • All prices are in EURO excl. VAT and shipping. Only available in selected countries.